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MB Sample ID: SA053328
Local Sample ID: | S43 |
Subject ID: | SU000947 |
Subject Type: | Sheep |
Subject Species: | Ovis aries |
Taxonomy ID: | 9940 |
Gender: | Both |
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Sample Preparation:
Sampleprep ID: | SP000954 |
Sampleprep Summary: | After sample randomization, weighed tissue (around 10mg for adipose and 50mg for arteries) in 2 mL polypropylene Eppendorf tube was enriched with deuterated surrogates in 20µL methanol (Tables S1 and S2 from Agrawal, K., L.A. Hassoun, N. Foolad, T.L. Pedersen, R.K. Sivamani, J.W. Newman. 2017. Sweat lipid mediator profiling: a non-invasive approach for cutaneous research. J. Lipid Res. 58:188–195 [EPub: Nov 7, 2016]. doi: 10.1194/jlr.M071738) and 5 μl of BHT/EDTA in 1:1 methanol/water (v/v). A total of 400 μl 1-cyclohexyl uredio, 3-dodecanoic acid / 1-phenyl ureido, 3-hexanoic acid (CUDA / PUHA) in 1:1 methanol/acetonitrile (v/v) was added, the sample ground with 3mm stainless steel beads (two for adipose tissue and 3 for blood vessel tissue) and agitation for 6 min at 1500RPM. Protein precipitate and debris were removed by centrifugation (10 min, 10,000 RCF, 6 °C). The supernatant was filtered by centrifugation through 1 µm PVDF membranes (Millipore, Billerica, MA) at 6 °C and 4,500 RCF for 3 min. The filtrate was stored in glass vials at -20 °C until UPLC-MS/MS analysis. |